Team:Warsaw/Calendar-Main/18 September 2008
From 2008.igem.org
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<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_purification_after_enzymatic_reaction">Clean-up</a> of digested PCR product.</li> | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_purification_after_enzymatic_reaction">Clean-up</a> of digested PCR product.</li> | ||
- | <li> Gel electrophoresis of digested plasmid and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band (<a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> - 2200 bp).</li></ol> | + | <li> Gel electrophoresis of digested plasmid and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper band (<a href=http://partsregistry.org/wiki/index.php?title=Part:pSB1A3>pSB1A3</a> - 2200 bp).<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/18_September_2008#fig3">Fig. 3</a>.</li></ol> |
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<a name="fig2"><img src="https://static.igem.org/mediawiki/2008/d/d0/Go_17_09.jpg" width=300/></a><var><b>Fig. 2. PCR to obtain inserts</b><br> | <a name="fig2"><img src="https://static.igem.org/mediawiki/2008/d/d0/Go_17_09.jpg" width=300/></a><var><b>Fig. 2. PCR to obtain inserts</b><br> | ||
1. Marker<br> | 1. Marker<br> | ||
- | 2. | + | 2. DeltaA<br> |
- | 3. | + | 3. Omega<br> |
- | 4. | + | 4. OmpA_omega<br></var> |
</p> | </p> | ||
+ | <p class="hide"><br> | ||
+ | <a name="fig3"><img src="https://static.igem.org/mediawiki/2008/9/96/Go2_19_09.jpg" width=300/></a><var><b>Fig. 3.</b> EcoRI/BcuI digest of pSB1A3 <br> | ||
+ | 1. Marker<br> | ||
+ | 2. Digested pSB1A3<br></var> | ||
+ | </p> | ||
+ | |||
+ | |||
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primers (annealing temperature 65 °C; elongation length 60s) to obtain <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103009>linker_alpha (BBa_K103009) fragment. </li> | primers (annealing temperature 65 °C; elongation length 60s) to obtain <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103009>linker_alpha (BBa_K103009) fragment. </li> | ||
- | <li> Gel electrophoresis of PCR product | + | <li> Gel electrophoresis of PCR product and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper bands (linker_alpha - 600 bp).<a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/18_September_2008#fig5">Fig. 5</a>.</li> |
<li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of purified PCR product with EcoRI and BcuI (BamHI buffer). </li> | <li><a href="https://2008.igem.org/Wiki/Team:Warsaw/igem_notebook.htm#digest">Digest</a> of purified PCR product with EcoRI and BcuI (BamHI buffer). </li> | ||
<li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_purification_after_enzymatic_reaction">Clean-up</a> of digested PCR product.</li> | <li><a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_purification_after_enzymatic_reaction">Clean-up</a> of digested PCR product.</li> | ||
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<a name="fig2"><img src="https://static.igem.org/mediawiki/2008/d/d0/Go_17_09.jpg" width=300/></a><var><b>Fig. 2.</b> PCR to obtain inserts<br> | <a name="fig2"><img src="https://static.igem.org/mediawiki/2008/d/d0/Go_17_09.jpg" width=300/></a><var><b>Fig. 2.</b> PCR to obtain inserts<br> | ||
1. Marker<br> | 1. Marker<br> | ||
- | 2. | + | 2. DeltaA<br> |
- | 3. | + | 3. Omega<br> |
- | 4. | + | 4. OmpA_omega<br></var> |
</p> | </p> | ||
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<a name="fig2"><img src="https://static.igem.org/mediawiki/2008/d/d0/Go_17_09.jpg" width=300/></a><var><b>Fig. 2.</b> PCR to obtain inserts<br> | <a name="fig2"><img src="https://static.igem.org/mediawiki/2008/d/d0/Go_17_09.jpg" width=300/></a><var><b>Fig. 2.</b> PCR to obtain inserts<br> | ||
1. Marker<br> | 1. Marker<br> | ||
- | 2. | + | 2. DeltaA<br> |
- | 3. | + | 3. Omega<br> |
- | 4. | + | 4. OmpA_omega<br></var> |
</p> | </p> | ||
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- | + | ||
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4. ompA_omega<br></var> | 4. ompA_omega<br></var> | ||
- | <a name="fig3"><img src="https://static.igem.org/mediawiki/2008/d/d0/Go_17_09.jpg" width=300/></a><var><b>Fig. | + | <a name="fig3"><img src="https://static.igem.org/mediawiki/2008/9/96/Go2_19_09.jpg" width=300/></a><var><b>Fig. 3.</b> EcoRI/BcuI digest of pSB1A3 <br> |
+ | 1. Marker<br> | ||
+ | 2. Digested pSB1A3<br></var> | ||
+ | |||
+ | <a name="fig5"><img src="https://static.igem.org/mediawiki/2008/d/d0/Go_17_09.jpg" width=300/></a><var><b>Fig. 5. PCR to obtain linker_alpha</b><br> | ||
1. Marker<br> | 1. Marker<br> | ||
2. linker_alpha<br></var> | 2. linker_alpha<br></var> | ||
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2. digested pSB1A3<br></var> | 2. digested pSB1A3<br></var> | ||
- | + | ||
- | + | ||
- | + | ||
</html> | </html> |
Revision as of 13:22, 27 October 2008
'Hunter and prey' system tests: Competition testsWeronikaInoculation of pACYC177+OmpA_A_alpha, pACYC177+OmpA_Z_omega, pACYC177+OmpA_A_omega and pACYC177+OmpA_Z_alpha + induction using 0.25mM iPTG MutD5 testingPiotrInoculation of mutD5 into 100 ml of LB and rendering them chemocompetent. Electroporation with pACYC177+OmpA_Z_alpha, pACYC177+OmpA_Z_omega, pACYC177+OmpA_A_omega and pACYC177+OmpA_omega_ΔA. Optimisation of primers for preparation of partsMichał K.
1. Marker 2. 55 °C linker_alpha 3. 60 °C linker_alpha 4. 65 °C linker_alpha 5. 70 °C linker_alpha Preparation of ΔA (BBa_K103003)Michał K.
Preparation of linker_alpha (BBa_K103009)Michał K.
Preparation of linker_omega (BBa_K103013)Michał K.
Preparation of OmpA-linker-omega-linker (BBa_K103016)Michał K.
Preparation of OmpA-linker (BBa_K103006)Michał K.
1. Marker 2. deltaA 3. omega 4. ompA_omega Fig. 3. EcoRI/BcuI digest of pSB1A3 1. Marker 2. Digested pSB1A3 Fig. 5. PCR to obtain linker_alpha 1. Marker 2. linker_alpha Fig. 4. Control SacI/NdeI digest of pET15b_OmpA_omega 1. Marker 2. digested pSB1A3
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